肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域研究

doi:10.12007/j.issn.0258-4646.2018.08.002

中国医科大学学报 ›› 2018, Vol. 47 ›› Issue (8): 678-681.doi: 10.12007/j.issn.0258-4646.2018.08.002

肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域研究

韩旭, 于潜, 田野, 赵希彤, 张磊, 周镝, 田大力

中国医科大学附属第四医院胸外科, 沈阳 110032

收稿日期:2018-01-22 出版日期:2018-08-30 发布日期:2018-08-17
通讯作者: 田大力 E-mail:cmu4h_dlt@hotmail.com
作者简介:韩旭(1979-),男,主治医师,博士.
基金资助:
国家自然科学基金（30973502，30170914）；辽宁省教育厅科学研究一般项目（L2013302）

Interacting Domain between ABCE1-encoding Protein and the Cytoskeleton Protein Actin

HAN Xu, YU Qian, TIAN Ye, ZHAO Xitong, ZHANG Lei, ZHOU Di, TIAN Dali

Department of Thoracic Surgery, The Fourth Affiliated Hospital, China Medical University, Shenyang 110032, China

Received:2018-01-22 Online:2018-08-30 Published:2018-08-17

摘要/Abstract

摘要： 目的明确肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域。方法通过转化技术，在E.coli BL21（DE3）中诱导表达前期研究获得的表达载体pGEX-4T-1-ABCE1-55，获得GST-ABCE1-55重组融合蛋白表达，应用GST pull-down的方法验证pGEX-4T-1-ABCE1-55与β-肌动蛋白有无相互作用。结果 GST-ABCE1-55是带有GST标签的含有357个氨基酸的重组蛋白，大小约66×10³，在GST pull-down实验中，GST-ABCE1-55并不能与β-肌动蛋白特异结合。结论肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域应位于ABCE1蛋白第358位至第600位编码氨基酸中。

关键词: ATP结合盒转运子E1, pGEX-4T-1原核表达载体, 重组蛋白表达, 蛋白质相互作用结构域

Abstract: Objective To investigate the interacting domain between ABCE1-encoding protein and the cytoskeleton protein actin. Methods Escherichia coli cells were induced with IPTG to produce the recombinant protein glutathione S-transferase(GST)-ABCE1-55 using the prokaryotic expression vector pGEX-4T-1-ABCE1-55,which was previously acquired. A GST pull-down assay was conducted to verify the interaction between GST-ABCE1-55 and β-actin. Results The recombinant protein GST-ABCE1-55 contained a GST tag and was 66×10³. There was no interaction between GST-ABCE1-55 and β-actin. Conclusion The interacting domain was predicted to be present between the 358th and 600th encoded amino acids.

Key words: ATP-binding cassette E1, prokaryotic expression vector pGEX-4T-1-ABCE1, recombinant protein expression, interacting domain

中图分类号:

R734.2

韩旭, 于潜, 田野, 赵希彤, 张磊, 周镝, 田大力. 肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域研究[J]. 中国医科大学学报, 2018, 47(8): 678-681.

HAN Xu, YU Qian, TIAN Ye, ZHAO Xitong, ZHANG Lei, ZHOU Di, TIAN Dali. Interacting Domain between ABCE1-encoding Protein and the Cytoskeleton Protein Actin[J]. Journal of China Medical University, 2018, 47(8): 678-681.

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中国医科大学学报

肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域研究

Interacting Domain between ABCE1-encoding Protein and the Cytoskeleton Protein Actin

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